EP3542826A1 - Nanoporteur pour le marquage fluorescent sélectif des cellules cancéreuses, et procédé de préparation de ce dernier - Google Patents

Nanoporteur pour le marquage fluorescent sélectif des cellules cancéreuses, et procédé de préparation de ce dernier Download PDF

Info

Publication number
EP3542826A1
EP3542826A1 EP17871175.0A EP17871175A EP3542826A1 EP 3542826 A1 EP3542826 A1 EP 3542826A1 EP 17871175 A EP17871175 A EP 17871175A EP 3542826 A1 EP3542826 A1 EP 3542826A1
Authority
EP
European Patent Office
Prior art keywords
nanocarrier
oil
cancer cell
phase ingredient
cancer
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP17871175.0A
Other languages
German (de)
English (en)
Other versions
EP3542826A4 (fr
Inventor
Kang Won Lee
Yoon Jeong
Sara Lee
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
SNU R&DB Foundation
Original Assignee
Seoul National University R&DB Foundation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Seoul National University R&DB Foundation filed Critical Seoul National University R&DB Foundation
Publication of EP3542826A1 publication Critical patent/EP3542826A1/fr
Publication of EP3542826A4 publication Critical patent/EP3542826A4/fr
Pending legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
    • A61K41/0061—5-aminolevulinic acid-based PDT: 5-ALA-PDT involving porphyrins or precursors of protoporphyrins generated in vivo from 5-ALA
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
    • A61K47/551—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6905—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion
    • A61K47/6907—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion the form being a microemulsion, nanoemulsion or micelle
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A61K49/0036—Porphyrins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
    • A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
    • A61K49/0076—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form dispersion, suspension, e.g. particles in a liquid, colloid, emulsion
    • A61K49/0082—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form dispersion, suspension, e.g. particles in a liquid, colloid, emulsion micelle, e.g. phospholipidic micelle and polymeric micelle
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1806—Suspensions, emulsions, colloids, dispersions
    • A61K49/1809—Micelles, e.g. phospholipidic or polymeric micelles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G01N33/57496—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving intracellular compounds
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
    • G01N33/587—Nanoparticles

Definitions

  • 5-Aminolevulinic acid has been used as a fluorescent substance for tumor surgery since 1979, and is a substance having few side effects when used clinically.
  • 5-ALA is known to produce protoporphyrin IX (PpIX) by reacting with the patient's glioma cells.
  • PpIX protoporphyrin IX
  • 5-ALA is converted into PpIX which is an intermediate in the heme biosynthesis pathway within mitochondria.
  • 5-ALA in itself has no fluorescent properties, but PpIX produced by reacting 5-ALA with cancer cells emits fluorescence of 635 nm at an excitation wavelength of about 400 nm to distinguish malignant glioma from normal tissues.
  • contrast agents including 5-ALA which are used during optical diagnosis and surgery are non-specific to a lesion, accurate diagnosis and surgery are difficult. Therefore, to impart target specificity to contrast agents, a method of crosslinking contrast agents with a tumor-specific ligand such as a lesion-specific peptide, antibody, or polysaccharide via a covalent bond has been actively employed.
  • a tumor-specific ligand such as a lesion-specific peptide, antibody, or polysaccharide via a covalent bond
  • new problems arise, such as reduction in chemical structure stability and targetability of the complex, and side effects in the human body, and thus accurate diagnosis and surgical resection of cancer are difficult.
  • biotin's own targetability may be reduced due to the covalent binding of the fluorescence-inducing substance to the targeting moiety, and efficacy stability may be reduced due to the structural change of biotin.
  • An aspect of the present disclosure provides a drug carrier for a contrast agent which may be used in cancer diagnosis, the drug carrier capable of targeting cancer cells without using a covalent bond with a tumor-specific ligand.
  • Another aspect of the present disclosure provides medical use of the drug carrier in cancer diagnosis.
  • Still another aspect of the present disclosure provides a method of preparing the drug carrier.
  • An aspect of the present disclosure provides a micelle structured nanocarrier including an aqueous phase ingredient, the aqueous phase ingredient obtained by dispersing, in water, a water-in-oil nanoemulsion including an oil phase ingredient, a surfactant, and the aqueous phase ingredient to remove the oil phase ingredient, wherein the aqueous phase ingredient includes a cancer cell fluorescence-inducing substance and a cancer cell-targeting polysaccharide.
  • Another aspect of the present disclosure provides a pharmaceutical composition for cancer diagnosis, the pharmaceutical composition including the micelle structured nanocarrier according to an aspect of the present disclosure.
  • Still another aspect of the present disclosure provides a method of preparing the micelle structured nanocarrier according to an aspect of the present disclosure.
  • the nanocarrier since the nanocarrier has nanoparticle size homogeneity and excellent thermodynamic stability, and its long-term storage is also possible, the nanocarrier is a pharmaceutically superior agent. Furthermore, since the nanocarrier may be prepared in a relatively simple manner and its mass production is possible, the nanocarrier is economical.
  • the present inventors have studied a drug carrier capable of targeting cancer cells without using a covalent bond between a cancer cell fluorescence-inducing substance such as 5-ALA and a tumor-specific ligand, and as a result, they found that when a nanocarrier is prepared by including both of a cancer cell fluorescence-inducing substance and a cancer cell-targeting polysaccharide inside an aqueous phase of the nanocarrier, the cancer cell fluorescence-inducing substance is selectively internalized into cancer cells, thereby identifying a cancerous tissue by fluorescence.
  • a cancer cell fluorescence-inducing substance such as 5-ALA and a tumor-specific ligand
  • the nanocarrier may be obtained by mixing an oil phase ingredient; a surfactant; and the aqueous phase ingredient including the cancer cell fluorescence-inducing substance and the cancer cell-targeting polysaccharide to prepare a water-in-oil nanoemulsion, and then dispersing the nanoemulsion in water to remove an oil phase ingredient.
  • an aspect of the present disclosure provides a micelle structured nanocarrier including an aqueous phase ingredient, the aqueous phase ingredient obtained by dispersing, in water, a water-in-oil nanoemulsion including an oil phase ingredient, a surfactant, and the aqueous phase ingredient to remove the oil phase ingredient, wherein the aqueous phase ingredient includes a cancer cell fluorescence-inducing substance and a cancer cell-targeting polysaccharide.
  • the nanocarrier is a micelle structured nanocarrier that includes the aqueous phase including the cancer cell fluorescence-inducing substance and the cancer cell-targeting polysaccharide inside thereof and the surfactant on the surface thereof.
  • oil phase ingredient refers to an oil-soluble substance that is dissolved in oil.
  • the oil phase ingredient may be any oil which may be used in the art for the preparation of the nanoemulsion, and for example, may be an oil selected from the group consisting of soybean oil, olive oil, grape seed oil, canola oil, corn oil, mineral oil, silicone oil, castor oil, paraffin oil, and any combination thereof.
  • the oil phase ingredient may be soybean oil.
  • cancer cell fluorescence-inducing substance refers to any substance that is internalized into cancer cells in vivo to generate a fluorescent substance.
  • the cancer cell fluorescence-inducing substance may be any substance that is known in the art to be internalized into cancer cells to generate a fluorescent substance or may be found in the future.
  • a substance capable of generating a fluorescent substance such as protoporphyrin IX may be a cancer cell fluorescence-inducing substance selected from the group consisting of heme, hemin, zinc protoporphyrin, magnesium protoporphyrin, hematoporphyrin, benzoporphyrin, metalloporphyrin, 5-aminolevulinic acid, texaphyrins, chlorins, purpurins, bacteriochlorins, phthalocyanine, naphthalocyanine, and derivatives thereof, and any combination thereof, but is not limited thereto.
  • the cancer cell fluorescence-inducing substance is 5-ALA.
  • cancer cell-targeting polysaccharide refers to any polysaccharide capable of selectively binding to a molecule or a receptor overexpressed on the surface of cancer cells.
  • the cancer cell-targeting polysaccharide may be any cancer cell-targeting polysaccharide that is known in the art to selectively bind to a molecule or a receptor overexpressed on the surface of cancer cells or may be found in the future.
  • cancer cell-targeting ligand-binding polysaccharide refers to a complex of a polysaccharide and any cancer cell-targeting ligand capable of selectively binding to a marker, i.e., a molecule or a receptor overexpressed on the surface of cancer cells via a covalent bond.
  • a marker i.e., a molecule or a receptor overexpressed on the surface of cancer cells via a covalent bond.
  • the marker overexpressed on the surface of cancer cells, the cancer cell-targeting ligand capable of binding thereto, and a method of binding the cancer cell-targeting ligand to the polysaccharide are well known in the art.
  • the cancer cell-targeting polysaccharide may be hyaluronic acid.
  • the hyaluronic acid may bind to CD44 which is a hyaluronic acid receptor specifically overexpressed on cancer cells, and thus hyaluronic acid serves as the cancer cell-targeting polysaccharide.
  • CD44 is a hyaluronic acid receptor specifically overexpressed on cancer cells
  • hyaluronic acid serves as the cancer cell-targeting polysaccharide.
  • most natural polysaccharides such as alginic acid, chitosan, pectin, etc. are known to have no specific cancer cell-binding ability, but these polysaccharides may be provided with targetability by covalently binding to the cancer cell-targeting ligand.
  • the cancer cell-targeting ligand-binding polysaccharide may bind to a receptor specifically overexpressed on the surface of cancer cells, wherein the receptor is able to bind with the ligand, and thus it may serve as a cancer cell-targeting polysaccharide.
  • the aptamer may be an aptamer binding to mucin, particularly mucin 1 (Muc1).
  • Mucin is an internal transmembrane domain which is a cell surface-associated glycoprotein attached to the cell.
  • mucin 1 (Muc1) contains a hydrophobic membrane-spanning domain with 31 amino acids, a cytoplasmic domain with 69 amino acids, and an extracellular domain consisting of nearly identical repeats with 20 amino acids. Muc1 is over-expressed in almost all human epithelial cancer cells including breast cancer, stomach cancer, colorectal cancer, lung cancer, prostate cancer, ovarian cancer, pancreatic cancer, and bladder cancer.
  • MUC1 the expression of MUC1 in these tissues lacks otherwise regular expression patterns, resulting in a ubiquitous, random expression of the protein all over the cell surface. Therefore, an aptamer (Anti-MUC1 Aptamer) specifically binding to mucin 1 (MUC1) may serve as the cancer cell-targeting ligand, and a polysaccharide binding thereto may be used as the cancer cell-targeting polysaccharide.
  • an aptamer specifically binding to mucin 1 (MUC1) specifically binding to mucin 1 (MUC1) may serve as the cancer cell-targeting ligand, and a polysaccharide binding thereto may be used as the cancer cell-targeting polysaccharide.
  • the micelle structured nanocarrier may have nanoparticles of an interpenetrating polymer network structure (IPN).
  • IPN interpenetrating polymer network structure
  • the interpenetrating polymer network structure refers to an entangled network formed by two or more components without a covalent bond.
  • FIG. 1(B) An illustration of an interpenetrating polymer network structure of a nanocarrier according to an embodiment of the present disclosure is shown in FIG. 1(B) .
  • the micelle structured nanocarrier may have the interpenetrating polymer network structure, the cancer cell fluorescence-inducing substance and the cancer cell-targeting polysaccharide included in the aqueous phase ingredient thereof may be physically encapsulated, thereby increasing mechanical strength and thermodynamic stability.
  • the nanocarrier may have a high absolute value of a zeta potential on the surface due to cations or anions of the cancer cell-targeting polysaccharide.
  • the zeta potential of the micelle structured nanocarrier may be -10 mV to -50 mV or 10 mV to 50 mV, and more specifically -10 mV to -30 mV or 10 mV to 30 mV.
  • the zeta potential is a value obtained when the cancer cell-targeting polysaccharide is negatively charged in the aqueous phase.
  • the surface zeta potential value is changed by an ionic bond by an interaction between the substances encapsulated inside the nanocarrier.
  • the nanoparticle may have an average particle size of 200 nm or less, and a zeta potential value of -10 mV to -30 mV. In terms of homogeneity of the nanoparticles, the nanoparticle may have an average particle size of about 100 nm, and a zeta potential value of -10 mV to -30 mV.
  • the cosurfactant may be a mixture containing sorbitan fatty acid ester (Span) and polyoxyethylene sorbitan fatty acid ester (Tween) and having the HLB value of 6 to 9.
  • the surfactant may be a cosurfactant containing Tween 80 (HLB 15) and Span 80 (HLB 4.3) and having the HLB value of about 7.
  • Another aspect of the present disclosure provides a pharmaceutical composition for cancer diagnosis, the pharmaceutical composition including the nanocarrier according to an aspect of the present disclosure.
  • the above description of the micelle structured nanocarrier according to an aspect of the present disclosure may be also applied to a detailed description of the nanocarrier included in the pharmaceutical composition for cancer diagnosis.
  • FIG. 1 illustrates a micelle structured nanocarrier containing 5-ALA and hyaluronic acid inside thereof according to an embodiment of the present disclosure (A), an interpenetrating polymer network structure of the nanocarrier (B), no response of the nanocarrier to a normal cell when it encounters the normal cell and a cancer cell (C), and a cellular interaction between the hyaluronic acid protruding from the nanocarrier and CD44 receptor on the surface of cancer cell (D).
  • Hyaluronic acid is encapsulated in the nanocarrier, but a portion of hyaluronic acid protrudes from the surface of the nanocarrier due to its chain-shaped molecular structure.
  • the micelle structured nanocarrier may selectively bind to cancer cells by binding to CD44 receptor overexpressed on the surface of cancer cell through the hyaluronic acid encapsulated in the nanocarrier.
  • the binding between the hyaluronic acid and CD44 receptor is attributed to a specific interaction between the chain structure present in hyaluronic acid and CD44 receptor overexpressed on the cancer cell.
  • 5-ALA in the nanocarrier binding to the surface of cancer cell may be internalized via CD44 receptor-mediated endocytosis.
  • the micelle structured nanocarrier does not generate fluorescence before internalization of 5-ALA into cancer cells.
  • 5-ALA is internalized into cancer cells by the targeting effect of hyaluronic acid
  • 5-ALA is induced to protoporphyrin IX by mitochondria inside the cytoplasm of cancer cells, and the produced protoporphyrin IX emits fluorescence of 635 nm when irradiated with light of a wavelength of 410 nm.
  • Selective uptake of the micelle structured nanocarrier according to an embodiment by cancer cells, and emitting of fluorescence thereby are shown in FIG. 2 .
  • the micelle structured nanocarrier according to an embodiment of the present disclosure may clearly distinguish cancer cells from a normal tissue by fluorescence, thereby being effectively used in cancer diagnosis.
  • the nanocarrier according to an embodiment of the present disclosure does not generate fluorescence with respect to normal cells, whereas selectively generates fluorescence with respect to cancer cells.
  • the micelle structured nanocarrier including the aqueous phase including hyaluronic acid and 5-ALA prepared according to Example 1 and a nanocarrier (Comparative Example 1) including, instead of hyaluronic acid, alginic acid which is not a cancer cell-targeting polysaccharide were treated to 4 different kinds of cells (fibroblast, glioma, lung carcinoma, and gastric adenocarcinoma).
  • the alginic acid-based nanocarrier did not generate selective fluorescence with respect to cancer cells, whereas the hyaluronic acid-based nanocarrier generated selective fluorescence with respect to cancer cells ( FIGS. 10 to 13 ).
  • composition for cancer diagnosis includes all of those used as a contrast agent to diagnose the presence of cancer as well as to monitor the treatment response or the severity of cancer during cancer therapy. Further, it is a concept including use of the contrast agent to clearly distinguish a cancerous tissue from a normal tissue during surgical resection of the cancerous tissue. In addition, it is construed to include any beneficial application that may be obtained by distinguishing a cancerous tissue from a normal tissue by fluorescence.
  • the cancer may be any cancer which may be targeted by the cancer cell-targeting polysaccharide and where the fluorescent substance may be induced from the cancer cell fluorescence-inducing substance, and may differ depending on the specific kind of the cancer cell fluorescence-inducing substance and/or the cancer cell-targeting polysaccharide.
  • the cancer may include brain tumor, lung cancer, stomach cancer, and ovarian cancer, but is not limited thereto.
  • the micelle structured nanocarrier is a nanocarrier showing no significant toxicity when administered into a living body.
  • the nanocarrier may include, as its raw materials, the cancer cell-targeting polysaccharide including a biocompatible polymer such as hyaluronic acid, alginic acid, or chitosan, etc. which has excellent biocompatibility, and 5-aminolevulinic acid as the cancer cell fluorescence-inducing substance which is also a non-toxic substance already present in a living body, and therefore, the nanocarrier may be used safely.
  • the cancer cell-targeting polysaccharide including a biocompatible polymer such as hyaluronic acid, alginic acid, or chitosan, etc. which has excellent biocompatibility
  • 5-aminolevulinic acid as the cancer cell fluorescence-inducing substance which is also a non-toxic substance already present in a living body, and therefore, the nanocarrier may be used safely.
  • the nanocarrier according to an embodiment of the present disclosure was used to perform a cytotoxicity test for fibroblast and three kinds of cancer cells (glioma, lung carcinoma, and gastric adenocarcinoma) by CCK-8 assay, and the results are shown in FIGS. 6 to 9 .
  • a cell viability test of the four cell groups showed that the nanocarrier had no toxicity even at a concentration of 2 mg/mL.
  • An administration dose of the composition for cancer diagnosis may vary depending on the kind of the cancer cell fluorescence-inducing substance, and in a specific embodiment, when the cancer cell fluorescence-inducing substance is 5-ALA, the component may be administered in an amount of about 0.1 mg to about 1,000 mg for an adult male, and the dose may be appropriately increased or decreased by a physician according to race, sex, age, body weight, kind of carcinoma, and the like.
  • the pharmaceutical composition for cancer diagnosis may be prepared in any formulation capable of delivering the nanocarrier to a cancerous tissue for cancer diagnosis, and for example, may be prepared in an injectable formulation.
  • a non-toxic buffer solution isotonic to blood may be included as a diluent, and for example, a phosphate buffer solution of pH 7.4 may be used.
  • the pharmaceutical composition may include other diluents or additives in addition to the buffer solution.
  • the excipients and additives which may be added to the injectable formulation are widely known to those skilled in the art, and for example, the following literature may be served as a reference ( Remington's Pharmaceutical Sciences (19th ed., 1995 ); Dr. H.P. Fiedler "Lexikon der Hilfsstoffe fur Pharmazie, Kosmetik und angrenzende füre" [Encyclopaedia of auxiliaries for pharmacy, cosmetics and related fields ]).
  • Still another aspect of the present disclosure provides a method of preparing the micelle structured nanocarrier according to an aspect of the present disclosure, the method including preparing the oil phase ingredient; preparing the surfactant; preparing the aqueous phase ingredient; mixing the oil phase ingredient, the surfactant, and the aqueous phase ingredient with stirring to prepare the water-in-oil nanoemulsion; and re-dispersing the water-in-oil nanoemulsion in water to remove an oil phase, thereby separating nanoparticles.
  • the above description of the micelle structured nanocarrier according to an aspect of the present disclosure may be also applied to a detailed description of the preparation method.
  • the preparing of the aqueous phase ingredient may include preparing a first aqueous phase ingredient including the cancer cell-targeting polysaccharide; and preparing a second aqueous phase ingredient including the cancer cell fluorescence-inducing substance.
  • the mixing with stirring may be performed by sonication, and according to a specific embodiment, high energy sonication may be performed for 5 minutes to 10 minutes using a 6 mm probe tip sonicator (Sonics, VC-750 amplitude 20-40%).
  • the preparation method may further include filtering, through a cellulose acetate syringe filter, the micelle structured nanocarrier which is obtained by removing the oil phase ingredient. Through the filtering, the nanocarrier may be obtained in a dispersed form without aggregation.
  • Span 80 (HLB value 4.3: Sigma) and Tween 80 (HLB value 15: Sigma) which are surfactants suitable for cell culture were prepared.
  • Span 80 and Tween 80 were used in combination to prepare a cosurfactant having an HLB value of 7.
  • a first aqueous phase ingredient (1 w/w% sodium alginate aqueous solution) and a second aqueous phase ingredient (5 w/w% 5-ALA aqueous solution) were prepared.
  • a mixed solution prepared by mixing the prepared oil phase ingredient (soybean oil): cosurfactant: first aqueous phase ingredient: second aqueous phase ingredient at a weight ratio of 7:1:1:1 was stirred by sonication (Sonic, VC-750 model, amplitude: 40%) for 10 minutes. The sonication was performed until the opaque mixture became transparent or translucent to prepare a water-in-oil (w/o) nanoemulsion.
  • a nanocarrier solution was prepared in the same manner as in Example 1, except that a 1%(w/w) 5-ALA aqueous solution was used instead of the 5%(w/w) 5-ALA aqueous solution as the second aqueous phase ingredient.
  • a nanocarrier solution was prepared in the same manner as in Example 1, except that a 1%(w/w) aqueous solution of folic acid covalently bound to sodium alginate (folic acid-alginic acid) was used instead of the 1%(w/w) sodium alginate aqueous solution.
  • a ratio of oil phase: cosurfactant: first aqueous phase: second aqueous phase used in the preparation of the nanoemulsions of Examples 1 to 4 are shown in Table 1 below.
  • Table 1 Weight ratio Oil phase Cosurfactant Aqueous phase Span80/Tween80
  • Example 1 (5-ALA NC1) 3.5 0.37/0.13 0.5 (1wt%HA)/ 0.5 (1wt% 5-ALA)
  • Example 2 (5-ALA NC2) 3.5 0.37/0.13 0.5 (1wt%HA)/ 0.5 (3wt% 5-ALA)
  • Example 3 (5-ALA NC3) 3.5 0.37/0.13 0.5 (1wt%HA)/ 0.5 (5wt% 5-ALA)
  • Example 4 (5-ALA NC4) 3.5 0.37/0.13 0.5 (1wt%FA-Alg)/ 0.5 (5wt% 5-ALA)
  • a nanocarrier solution was prepared in the same manner as in Example 1, except that a 1%(w/w) sodium alginate aqueous solution was used instead of the 1%(w/w) sodium hyaluronate aqueous solution.
  • Example 2 5 ⁇ l of the nanocarrier solution obtained in Example 1 was dropped on a 400-mesh copper grid of transmission electron microscope, followed by carefully wiping off around the grid with a towel. To remove excess water remaining in the grid, the grid was dried in a vacuum chamber for about 1 hour. After complete drying, the size of the nanocarrier was measured using a high magnification transmission electron microscope, and TEM images were photographed. The results are shown in FIG. 3 . In the left of FIG. 3 , TEM images of the nanocarrier which were observed at low and high magnifications are shown, and in the right of FIG. 3 , a graph of the particle size based on the image analysis is shown. It was found that an average particle size of the nanocarrier was about 53.58 nm, and a standard deviation was 16.25 nm.
  • Example 1 The average size and surface charge of the nanocarriers obtained in Examples 1 to 4 were measured using a dynamic light scattering (DLS) instrument (Zeta Nano ZS 3600, Malvern, UK). Each sample was measured in triplicate, and a mean value of the measured values was obtained. The results are shown in Table 2 below. [Table 2] Size (nm, DLS Z-avg) Surface charge (mV) Example 1 (5-ALA NC1) 88.72 ⁇ 1.5 -10.8 ⁇ 0.2 Example 2 (5-ALA NC2) 100.0 ⁇ 1.8 -12.9 ⁇ 0.1 Example 3 (5-ALA NC3) 144.1 ⁇ 2.6 -22.5 ⁇ 0.4 Example 4 (5-ALA NC4) 137.6 ⁇ 2.7 -10.7 ⁇ 0.3
  • the nanocarriers obtained in Examples 1 to 3 were refrigerated for 1 day to 60 days, and then an average particle size thereof was measured using a dynamic scattering light instrument. Each sample was measured in triplicate, and a mean value of the measured values was obtained. The results of measuring the average particle size of the nanocarriers over time are shown in FIG. 4 .
  • Cancer cells were treated with the nanocarrier obtained in Example 1, and it was examined whether protoporphyrin IX was actually induced in the cancer cells.
  • C6 rat glioma
  • A549 human lung carcinoma
  • MKN-74 human gastric adenocarcinoma
  • KCLB Korea Cell Line Bank
  • the three kinds of cancer cell groups were cultured in a 24-well plate at a density of 1 mL (5x10 5 cells/mL) for 24 hours, respectively. Then, the medium was replaced by a serum-free medium, and each of the cancer cells was treated with 100 ⁇ L (1 mg/mL concentration) of the nanocarrier obtained in Example 1, followed by incubation for 6 hours.
  • the cell culture medium was completely aspirated, and 100 ⁇ L of RIPA buffer solution was dropped to degrade cell membrane proteins.
  • Absorbance of the extracted protoporphyrin IX was measured ( FIG. 5B ), and fluorescence spectra ( FIG. 5C ) was measured using a spectrofluorometer. Fluorescence spectrum ( FIG. 5A ) of the nanocarrier itself before treatment to the cancer cells was also measured using a spectrofluorometer.
  • the result of fluorescence spectrum of the nanocarrier itself ( FIG. 5A ) showed no excitation wavelength and no emission wavelength between a wavelength of 350 nm and a wavelength of 750 nm, whereas the nanocarrier-treated cancer cells showed absorbance peak of protoporphyrin IX ( FIG. 5B ), suggesting that protoporphyrin IX was induced in the cancer cells, which was further supported by the fluorescence spectra of FIG. 5C .
  • the nanocarrier obtained in Example 1 was subjected to a cytotoxicity test.
  • the C6 (rat glioma), A549 (human lung carcinoma), and MKN-74 (human gastric adenocarcinoma) cell lines cultured in Experimental Example 4 were used.
  • 3T3-L1 (mouse fibroblast) cell line was also obtained from the Korea Cell Line Bank, and cultured in DEME (WELLGENE) supplemented with 1% penicillin-streptomycin (WELLGENE) and 10% bovine calf serum (BCS; WELLGENE) in an incubator at 37 °C, 5 % CO 2 , and subculturing was performed every three days.
  • the four kinds of cell lines were subjected to cell counting kit (CCK)-8 assay.
  • each of the four kinds of cells (5.0x10 4 /mL) was seeded in each well of a 96-well plate, and then cultured for 24 hours.
  • the medium was replaced by a serum-free medium, and then different concentrations (0.062 mg/mL, 0.125 mg/mL, 0.25 mg/mL, 0.5 mg/mL, 1 mg/mL, and 2 mg/mL) of the nanocarrier were added at a volume of 10% of the total volume.
  • 10 ⁇ g of CCK-8 solution (CCK-8; Dojindo) was added thereto, followed by further incubation for 2 hours.
  • Absorbance at 450 nm was measured using a microplate reader (iMark Bio-Rad instrumenis. Inc.). Absorbance was measured in triplicate under respective experimental conditions, and a mean value of the absorbance values was obtained, and compared with a mean value of absorbance values of a control group wherein a non-toxic phosphate buffer solution (PBS) was used instead of the nanocarrier to examine cytotoxicity.
  • PBS non-toxic phosphate buffer solution
  • each of the four kinds of cells (5.0x10 5 /mL) was seeded in each well of a 24-well plate, and then cultured for 24 hours.
  • the medium was replaced by a serum-free medium, and then 100 ⁇ L (1 mg/mL) of the nanocarrier corresponding to 10% of the total volume was added. After incubation for 1 hour, 3 hours, and 6 hours, cell fixation was performed. The medium in the well was completely aspirated, and then the well was was washed with PBS three times.
  • DAPI 4',6-diamidino-2-phenylindole staining was performed for cell nuclear staining.
  • Cells were stained with 1X DAPI for 3 minutes, and then washed with PBS three times.
  • cover glass was covered with DAPI and PplX fluorescence was examined using a fluorescence microscope (Carl Zeiss, Axiovert 200) under each condition.
  • the hyaluronic acid-based nanocarrier having cancer cell targetability showed PpIX fluorescence in all the cancer cells and no PpIX fluorescence in normal cells, whereas the alginic acid-based nanocarrier having no cancer cell targetability showed no PpIX fluorescence in all the cancer cells and normal cells.
  • DAPI fluorescence for nuclear staining was observed in all experimental groups, irrespective of the kind of the cancer cells and use of the hyaluronic acid-based nanocarrier.
  • Example 4 A cell uptake assay of the nanocarrier obtained in Example 4 which was based on folic acid-alginic acid as a cancer cell-targeting ligand-binding polysaccharide was performed.
  • A549 (human lung carcinoma) and SK-OV-3 (human ovarian carcinoma) cell lines were obtained from the Korea Cell Line Bank (KCLB) and cultured in RPMI 1640 (WELLGENE) supplemented with 1% penicillin-streptomycin (WELLGENE) and 10% fetal bovine serum (FBS; WELLGENE) in an incubator at 37 °C, 5 % CO 2 , and subculturing was performed every two days.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Cell Biology (AREA)
  • Dispersion Chemistry (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Analytical Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nanotechnology (AREA)
  • Hospice & Palliative Care (AREA)
  • Oncology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Radiology & Medical Imaging (AREA)
  • Medicinal Preparation (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
EP17871175.0A 2016-11-18 2017-02-20 Nanoporteur pour le marquage fluorescent sélectif des cellules cancéreuses, et procédé de préparation de ce dernier Pending EP3542826A4 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020160154441A KR101901986B1 (ko) 2016-11-18 2016-11-18 암세포의 선택적 형광 표지를 위한 나노전달체 및 그 제조방법
PCT/KR2017/001829 WO2018092984A1 (fr) 2016-11-18 2017-02-20 Nanoporteur pour le marquage fluorescent sélectif des cellules cancéreuses, et procédé de préparation de ce dernier

Publications (2)

Publication Number Publication Date
EP3542826A1 true EP3542826A1 (fr) 2019-09-25
EP3542826A4 EP3542826A4 (fr) 2020-08-05

Family

ID=62145544

Family Applications (1)

Application Number Title Priority Date Filing Date
EP17871175.0A Pending EP3542826A4 (fr) 2016-11-18 2017-02-20 Nanoporteur pour le marquage fluorescent sélectif des cellules cancéreuses, et procédé de préparation de ce dernier

Country Status (6)

Country Link
US (1) US11298428B2 (fr)
EP (1) EP3542826A4 (fr)
JP (1) JP6996774B2 (fr)
KR (1) KR101901986B1 (fr)
CN (1) CN110325221B (fr)
WO (1) WO2018092984A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102574572B1 (ko) 2021-02-02 2023-09-06 서울대학교 산학협력단 알긴산-엽산 컨쥬게이트의 제조방법, 이로부터 제조된 알긴산-엽산 컨쥬게이트 및 이를 포함하는 약학 조성물
CN115429899A (zh) * 2021-06-01 2022-12-06 牛尾电机(苏州)有限公司 试剂的制造方法及利用该制造方法制造的试剂

Family Cites Families (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR0178860B1 (ko) * 1996-08-08 1999-03-20 이서봉 에멀젼 네트웍을 이용한 국소 부위 약물 전달체계 및 그 제조 방법
SE507490C2 (sv) * 1996-08-27 1998-06-15 Medeikonos Ab Förfarande för detektering av cancer i hud på människor och däggdjur samt anordning för utövande av förfarandet
AU5135698A (en) 1996-11-28 1998-06-22 Nihon Schering K.K. Contrast compound, contrast medium for mri, and method for mri
NZ507540A (en) 1998-04-28 2002-07-26 Jagotec Ag Hyaluronic acid based imaging agents containing gadolinium or iron oxide
US20100062000A1 (en) * 2006-11-21 2010-03-11 The Regents Of The University Of California Rhamm, a Co-Receptor and Its Interactions with Other Receptors in Cancer Cell Motility and the Identification of Cancer Prognitor Cell Populations
KR100852944B1 (ko) * 2007-02-15 2008-08-19 (주)아모레퍼시픽 화학적으로 가교된 히알루론산 하이드로겔 나노입자의제조방법
JP5523338B2 (ja) * 2007-12-19 2014-06-18 エヴォニク ゴールドシュミット ゲーエムベーハー エマルジョン中の架橋ヒアルロン酸
FR2934955B1 (fr) * 2008-08-14 2011-07-08 Commissariat Energie Atomique Encapsulation d'agents therapeutiques lipophiles ou amphiphiles dans des nanoemulsions
US20100331819A1 (en) * 2009-06-24 2010-12-30 Abbott Cardiovascular Systems Inc. Drug Delivery System and Method of Treatment of Vascular Diseases Using Photodynamic Therapy
KR101309227B1 (ko) 2011-02-28 2013-09-23 부산대학교 산학협력단 광역동 치료에 사용되는 아미노레불리닉산이 공유결합된 히아루론산 나노입자
US9241921B2 (en) 2011-05-02 2016-01-26 Pankaj Modi Photosensitizer composition for treating skin disorders
CN102552174B (zh) * 2011-11-18 2015-07-22 中山大学 一种高载药量高包封率多肽/蛋白类药物纳米粒的制备方法
WO2013124444A1 (fr) * 2012-02-24 2013-08-29 INSERM (Institut National de la Santé et de la Recherche Médicale) Billes de polyssaccharide réticulé comprenant un agent d'imagerie
CN102697731B (zh) * 2012-05-10 2013-12-25 华东医院 5-氨基酮戊酸纳米粒及其制备方法和装置
KR101555764B1 (ko) * 2013-12-27 2015-10-01 연세대학교 산학협력단 고분자 하이드로겔 복합체 및 이의 제조방법
CN103976956B (zh) * 2014-05-16 2016-08-24 暨南大学 一种靶向抗肝癌纳米粒子及其制备方法和应用
CN105727305B (zh) * 2016-02-24 2018-09-07 温州医科大学 一种Legumain响应释放阿霉素缓释纳米制剂及制备方法与作为制备载体药物的应用
KR101939880B1 (ko) 2016-06-29 2019-01-18 서울대학교산학협력단 암병변 선택적 표지를 위한 수화젤 기반 나노 에멀전 및 이의 제조 방법

Also Published As

Publication number Publication date
KR20180056308A (ko) 2018-05-28
WO2018092984A1 (fr) 2018-05-24
JP2019537612A (ja) 2019-12-26
CN110325221B (zh) 2022-04-08
US11298428B2 (en) 2022-04-12
JP6996774B2 (ja) 2022-01-17
EP3542826A4 (fr) 2020-08-05
KR101901986B1 (ko) 2018-09-27
CN110325221A (zh) 2019-10-11
US20200268903A1 (en) 2020-08-27

Similar Documents

Publication Publication Date Title
Li et al. Facile strategy by hyaluronic acid functional carbon dot-doxorubicin nanoparticles for CD44 targeted drug delivery and enhanced breast cancer therapy
Peng et al. Herceptin-conjugated paclitaxel loaded PCL-PEG worm-like nanocrystal micelles for the combinatorial treatment of HER2-positive breast cancer
Choi et al. Self-assembled hyaluronic acid nanoparticles as a potential drug carrier for cancer therapy: synthesis, characterization, and in vivo biodistribution
Joseph et al. Galactoxyloglucan-modified nanocarriers of doxorubicin for improved tumor-targeted drug delivery with minimal toxicity
Zhang et al. Redox-and light-responsive alginate nanoparticles as effective drug carriers for combinational anticancer therapy
CN107669632B (zh) 药物载体、胶束、药物制剂、及其制备方法和用途
US20120087859A1 (en) Nanocarrier having enhanced skin permeability, cellular uptake and tumour delivery properties
Han et al. Redox-sensitive micelles for targeted intracellular delivery and combination chemotherapy of paclitaxel and all-trans-retinoid acid
WO2014155142A1 (fr) Nanocomposition stable comprenant de la doxorubicine, procédé pour la préparation de celle-ci, son utilisation et compositions pharmaceutiques contenant celle-ci
Vivek et al. Multifunctional nanoparticles for trimodal photodynamic therapy-mediated photothermal and chemotherapeutic effects
Oh et al. Radio-opaque theranostic nanoemulsions with synergistic anti-cancer activity of paclitaxel and Bcl-2 siRNA
Wang et al. Chitosan-and hyaluronic acid-based nanoarchitectures in phototherapy: Combination cancer chemotherapy, immunotherapy and gene therapy
Li et al. Temperature-and pH-responsive injectable chitosan hydrogels loaded with doxorubicin and curcumin as long-lasting release platforms for the treatment of solid tumors
Wang et al. Development and evaluation of hyaluronic acid-based polymeric micelles for targeted delivery of photosensitizer for photodynamic therapy in vitro
Le et al. Combined phototherapy with metabolic reprogramming-targeted albumin nanoparticles for treating breast cancer
Karimi-Soflou et al. Redox-Sensitive multifunctional hyaluronic acid-based nanomicelles with Fine-controlled anticancer drug release
EP2978428A1 (fr) Nanocomposition stable comprenant de l'épirubicine, procédé pour sa préparation, son utilisation et compositions pharmaceutiques la contenant
Zhao et al. Targeting delivery of partial VAR2CSA peptide guided N-2-Hydroxypropyl trimethyl ammonium chloride chitosan nanoparticles for multiple cancer types
US11298428B2 (en) Nanocarrier for selective fluorescence labeling of cancer cell and preparation method therefor
Guo et al. Hyaluronic acid/dextran-based polymeric micelles co-delivering ursolic acid and doxorubicin to mitochondria for potentiating chemotherapy in MDR cancer
US11103600B2 (en) Hydrogel-based nanoenulsion for selectively labeling cancer lesion, and preparation method therefor
Khan et al. Recent progress in Zein nanoparticles: A promising approach for advancing Cancer therapy
CN114848843A (zh) 一种化疗协同靶向联合治疗纳米药物及其在肿瘤治疗中的应用
KR102574572B1 (ko) 알긴산-엽산 컨쥬게이트의 제조방법, 이로부터 제조된 알긴산-엽산 컨쥬게이트 및 이를 포함하는 약학 조성물
Aravindan et al. Implications of Nanotherapeutic advancements to leverage Multi-Drug Resistant Breast Cancer: The State-of-the-Art Review

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20190618

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

AX Request for extension of the european patent

Extension state: BA ME

DAV Request for validation of the european patent (deleted)
DAX Request for extension of the european patent (deleted)
A4 Supplementary search report drawn up and despatched

Effective date: 20200707

RIC1 Information provided on ipc code assigned before grant

Ipc: G01N 33/58 20060101ALI20200701BHEP

Ipc: A61K 49/00 20060101AFI20200701BHEP

Ipc: A61K 49/18 20060101ALI20200701BHEP

Ipc: G01N 33/574 20060101ALI20200701BHEP